Immune adjuvant and application thereof

An immune adjuvant and vaccine technology, applied in antiviral agents, aerosol delivery, medical preparations containing active ingredients, etc., can solve the problems of poor immune regulation effect, achieve good adjuvant effect, and enhance different antigenic humors Effects of immune response and mucosal immune response

Inactive Publication Date: 2020-04-10
NAT VACCINE & SERUM INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] One aspect of the present invention is to provide a kind of immune adjuvant and its appl...

Method used

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  • Immune adjuvant and application thereof
  • Immune adjuvant and application thereof
  • Immune adjuvant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: Preparation of 1% chitosan and its derivative solution

[0052] Primary material sources:

[0053] Chitosan and chitosan acetate were purchased from HEPPE MEDICAL CHITOSAN GmbH

[0054] Chitosan quaternary ammonium salt was purchased from Nantong Lushen Biological Engineering Co., Ltd.

[0055] Methylene glycol chitosan was purchased from SIGMA

[0056] (1) Preparation of 1% chitosan quaternary ammonium salt (HTCC) solution

[0057] Accurately weighed 0.5 gram of chitosan quaternary ammonium salt is placed in a beaker with a magnetic stirrer, adds 50ml0.3% acetic acid solution, magnetic stirrer 200rpm stirs for 30 to 45 minutes, the chitosan quaternary ammonium salt after dissolving The ammonium salt solution was centrifuged at 4000rmp for 10 minutes, and finally the centrifuged supernatant was passed through a 0.45 μm filter membrane, and stored at 2-8°C for later use.

[0058] (2) Preparation of 1% methyl glycol chitosan (MGC) solution and chitosan a...

Embodiment 2

[0062] Example 2: Evaluation of the mucosal immune effect of chitosan derivatives as hepatitis B vaccine adjuvants

[0063]Chitosan and its derivatives in Example 1 are used as vaccine adjuvants, and recombinant hepatitis B subunit vaccine (Saccharomyces cerevisiae expresses HBsAg, produced by Beijing Tiantan Biological Products Co., Ltd.) is used as antigen. Take the preparation of 1ml of vaccine with chitosan or its derivatives as an example, take 100μl of chitosan or its derivatives solution, add 100μl of PBS and mix well, then add 200μl of normal saline, and finally add 550μl of HBsAg solution Mix quickly and store at 2-8°C. Used to immunize mice with intranasal drops, and detect anti-HBs in mouse serum, the specific method is as follows:

[0064] Experimental animals: BALB / c mice, 6-8 weeks old, 13 mice / group, female, purchased from Weitong Lihua Experimental Animal Technology Co., Ltd.

[0065] Drug concentration: chitosan and its derivatives prepared in Example 1 abov...

Embodiment 3

[0120] Embodiment 3: Chitosan derivative is used as the IgG of the mucous membrane immune adjuvant of influenza vaccine and hemagglutination inhibitory titer evaluation

[0121] Chitosan and derivatives thereof prepared in Example 1 were used as vaccine adjuvant, and H1N1 influenza vaccine (H1N1 influenza virus lysate, Beijing Tiantan Biological Products Co., Ltd.) was used as antigen. The two prepared vaccines according to the preparation method of hepatitis B vaccine in Example 2, immunized mice with nasal drops, and detected serum-specific IgG and hemagglutination inhibitory titers. The specific methods are as follows:

[0122] Experimental animals: BALB / c mice, 6-8 weeks old, 10 mice / group, female, purchased from Weitong Lihua Experimental Animal Technology Co., Ltd.

[0123] Drug concentration: Chitosan and its derivatives prepared in Example 1 above: 10 mg / ml; the final concentration of HA in the H1N1 lysate is 562 μg / ml.

[0124] Control solvent: normal saline (Normal ...

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Abstract

The invention discloses an immune adjuvant. The immune adjuvant comprises one or more selected from a salt chitosan solution, a quaternized chitosan solution or a hydroxylated chitosan solution, wherein the pH value of the salt chitosan solution is 4.5-5.5, the pH value of the quaternized chitosan solution is 5.8-8, and the pH value of the hydroxylated chitosan solution is 5.8-8. The screened specific chitosan derivatives can enhance humoral immune response and mucosal immune response of different antigens, have a good adjuvant effect, lay a foundation for research on the action mechanism of chitosan derivative adjuvants, and are expected to replace chitosan in development of mucosal adjuvants.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to an immune adjuvant and its application. Background technique [0002] Adjuvant (Adjuvant) is very important for the effect of vaccines. The use of immune adjuvants can sometimes greatly improve the protective effect of vaccines and enhance the specific immune response of the body's immune system to antigens. Currently approved human vaccine adjuvants include aluminum adjuvant, MF59 adjuvant, AS series adjuvant (AS04, AS03, AS01), virosome adjuvant, ISA51 adjuvant, CpG adjuvant, etc. [0003] Aluminum adjuvant is the first adjuvant approved for human vaccines in the world. Since it was first used in 1926, there have been 33 approved vaccines containing aluminum adjuvant in the United States, and at least 146 licensed vaccines worldwide The vaccine contains an aluminum adjuvant. The aluminum-containing compounds that have been used include aluminum hydroxide adjuvant, ...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/145A61K39/29A61K9/00A61K9/12A61P1/16A61P31/20A61P31/16
CPCA61K39/39A61K39/12A61K9/0043A61K9/12A61P1/16A61P31/20A61P31/16A61K2039/55583A61K2039/543C12N2730/10134C12N2760/16134
Inventor 张云涛徐静单璞高金爽邓霞王志标
Owner NAT VACCINE & SERUM INST
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